Characterization and protein engineering of glycosyltransferases for the biosynthesis of diverse hepatoprotective cycloartane-type saponins in Astragalus membranaceus

doi:10.1111/pbi.13983

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	Characterization and protein engineering of glycosyltransferases for the biosynthesis of diverse hepatoprotective cycloartane-type saponins in Astragalus membranaceus
	Chen,Kuan; Zhang,Meng; Gao,Baihan; Hasan,Aobulikasimu; Li,Junhao; Bao,Yang'oujie; Fan,Jingjing; Yu,Rong; Yi,Yang; Agren,Hans; Wang,Zilong; Liu,Haiyang; Ye,Min; Qiao,Xue
	2023
发表期刊	PLANT BIOTECHNOLOGY JOURNAL
ISSN	1467-7652
摘要	Although plant secondary metabolites are important source of new drugs, obtaining these compounds is challenging due to their high structural diversity and low abundance. The roots of Astragalus membranaceus are a popular herbal medicine worldwide. It contains a series of cycloartane-type saponins (astragalosides) as hepatoprotective and antivirus components. However, astragalosides exhibit complex sugar substitution patterns which hindered their purification and bioactivity investigation. In this work, glycosyltransferases (GT) from A. membranaceus were studied to synthesize structurally diverse astragalosides. Three new GTs, AmGT1/5 and AmGT9, were characterized as 3-O-glycosyltransferase and 25-O-glycosyltransferase of cycloastragenol respectively. AmGT1(G146V/I) variants were obtained as specific 3-O-xylosyltransferases by sequence alignment, molecular modelling and site-directed mutagenesis. A combinatorial synthesis system was established using AmGT1/5/9, AmGT1(G146V/S) and the reported AmGT8 and AmGT8(A394F). The system allowed the synthesis of 13 astragalosides in Astragalus root with conversion rates from 22.6% to 98.7%, covering most of the sugar-substitution patterns for astragalosides. In addition, AmGT1 exhibited remarkable sugar donor promiscuity to use 10 different donors, and was used to synthesize three novel astragalosides and ginsenosides. Glycosylation remarkably improved the hepatoprotective and SARS-CoV-2 inhibition activities for triterpenoids. This is one of the first attempts to produce a series of herbal constituents via combinatorial synthesis. The results provided new biocatalytic tools for saponin biosynthesis.
关键词	astragalosides biosynthesis glycosyltransferases protein engineering Astragalus membranaceus FUNCTIONAL-CHARACTERIZATION UDP-GLYCOSYLTRANSFERASES CARBON-TETRACHLORIDE PROTOPANAXATRIOL PHARMACOLOGY GENERATION CHEMISTRY ACCURACY PATHWAY
学科领域	Biotechnology & Applied Microbiology ; Plant Sciences
DOI	10.1111/pbi.13983
WOS记录号	WOS:000908340600001
引用统计
文献类型	期刊论文
条目标识符	http://ir.kib.ac.cn/handle/151853/74912
专题	中国科学院昆明植物研究所
推荐引用方式 GB/T 7714	Chen,Kuan,Zhang,Meng,Gao,Baihan,et al. Characterization and protein engineering of glycosyltransferases for the biosynthesis of diverse hepatoprotective cycloartane-type saponins in Astragalus membranaceus[J]. PLANT BIOTECHNOLOGY JOURNAL,2023.
APA	Chen,Kuan.,Zhang,Meng.,Gao,Baihan.,Hasan,Aobulikasimu.,Li,Junhao.,...&Qiao,Xue.(2023).Characterization and protein engineering of glycosyltransferases for the biosynthesis of diverse hepatoprotective cycloartane-type saponins in Astragalus membranaceus.PLANT BIOTECHNOLOGY JOURNAL.
MLA	Chen,Kuan,et al."Characterization and protein engineering of glycosyltransferases for the biosynthesis of diverse hepatoprotective cycloartane-type saponins in Astragalus membranaceus".PLANT BIOTECHNOLOGY JOURNAL (2023).

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